Therapeutic composition of a 5-aminohexahydro-pyrimidine and selected chelating agents



United States Patent I 3 072 529 THERAPEUTIC COMOSiTION OF A 5-MWIN- HEXAHYDRO PYRHVIIDINE This invention relates to certain novel compositions of matter and relates more particularly to broad spectrum antibacterial and antifungal compositions comprising the combination of achelating or sequestering agent with a S-amino-hexahydro-pyrimidine compound of the formula R-I\ N-R Hat i112 R NHZ where in R is an alkyl, aryl, aralkyl, alkaryl, hydroxyalkyl, aminoalkyl or cycloalkyl radical, and R is hydrogen, a lower alkyl or hydrcxymethyl radical. This invention is particularly concerned with the foregoing compositions wherein the chelating agents employed are those which are particularly adapted to remove or complex metals such as, for example, calcium, magnesium, iron, Zinc, manganese and cobalt.

Recent studies have indicated that the S-amino-hexahydro-pyrimidines having the general formula above are very useful antibacterial and antifungal agents. They are particularly valuable for topical application. Some indications of their value when they are employed systemically has also been observed. In using the S-amino-hexahydro-pyrimidines for topical application both on the skin and on the mucous membranes of various body cavities, it is preferred for economic and other reasons, that as low an active concentration as possible be used. Topically applied compositions containing concentrations above about 0.5% by Weight andespecially above about 1.0% by Weight have been found to be quite effective but in some instances, they have also been found to cause an undesirable degree of irritation. In addition, these higher concentrations frequently exert a local anesthetic act-ion which tends to limit the usefulness of these compositions in the higher concentrations.

It is therefore, an important object of this invention to provide effective antibacterial and antifungal compositions comprising a 5-amino-hexahydro-pyrimidine of the above structure which are of greatly enhanced potency and which are eifective both topically and systemically.

Another object of this invention is the preparation of antibacterial and antifungal compositions containing a 5-amino-hexahydro-pyrimidine wherein the effective concentration of the 5-amino-hexahydro-pyrimidine may be substantially decreased without any impairment in the antibacterial and antifungal potency of said compositions.

A further object of this invention is the provision of compositions containing a 5-amino-hexahydro-pyrimidine and a chelating or sequestering agent and which are highly active against a wide variety of microorganisms.

Other objects of this invention will appear from the following detailed description.

We have now found that the effective concentration of said 5-amino-hexahydro-pyramidines in antibacterialand antifungal compositions may be substantially reduced without affecting the potency of said compositions if a chelating agent such as, for example ethylenediamine tetraacetic acid, or any one or more of the several sodium and mixed sodium salts are employed in combination with the 5-amino-hexahydro-pyrimidine in said antibac- "ice terial and antifungal compositions. By formulating these compositions in the manner described, only a fraction of the amount of the 5-amino-hexahydro-pyrimidine usually employed to obtain the desired antibacterial and antifungal activity is required. The novel compositions of our invention are particularly valuable in that they are active in preventing the growth of gram-negative micrd organisms. This novel combination of active components is particularly effective in inhibiting and preventing the growth of the microorganism Ps. aeruginosa as well as actively inhibiting the growth of P. vulgaris, E. coli and C. albicans. While these microorganisms will exhibit growth in the presence of concentrations as high as 200 micrograms per milliliter of certain S-amino-hexahydropyrimidines, by the addition of as little as 0.1 to 10% on the weight of the composition of a chelating agent, for example 0.1% by Weight of ethylenediamine tetracetic acid, the growth of these organisms is completely inhibited in the presence of from only 1 to 10 micrograms per milliliter of the 5-amino-hexahydro-pyrimidine. Without impairing the marked activity of these compounds, the substantial reduction in concentration eliminates all possibility of irritation.

Examples of the 5amino-hexahydro-pyrimidines which may be employed in forming the novel compositions of our invention are:

1,3-bismethyl-S-methyl-S-amino hexahydro-pyrimidine 1,3-bis (beta-ethylhexyl)-5-methy1-S-amino-hexahydropyrimidine l,3-bispropyl-S-methyl-S-amino-hexahydro-pyrimidine l,3-bisisopropyl-5- nethyl-5-amino-hexahydro-pyrimidine 1,3-bis (secondary butyl)-5-methyl-5-amino-hexahydropyrimidine 1,3-bis (tertiary butyl)-5-methyl-5-amino-hexahydropyrimidine 1,3-bishexyl-5-methyl-5-amino-hexahydro-pyrimidine l,3-bisheptyl-S-methyl-5-amino-hexahydro-pyrimidine 1,3-bisoctyl-5-methyl-5-amino-hexahydro-pyrimidine l,3-bisdecyl-5-methyl-5-amino-hexahydro-pyrimidine 1,3-bisdodecyl-5-methyl-S-amino-hexahydro-pyrimidine 1,3-bistetradecyl-5-methyl-S-amino-hexahydropyrimidine 1,3-bishexadecyl-S-metl1yl-5-amino-hexahydropyrimidine 1,3-bis l-methylheptyl) -5-methyl-5-amino-hexahydropyrimidine 1,3-bis 1,3-dimethylbutyl) -5-methyl-5-amino-hexahydr0- pyrimidine 1,3-bis (tertiary butyl-Z-methyl)-5-methyl-5-aminohexahyidro-pyrimidine 1,3-bis (isopropyl-Z-methyl)-5-methyl-S-aminohexahydro-pyrirnidine 1,3-bis (hydroxy tertiary butyl)-5-methyl-5-aminohexahydro-pyrimidine 1,3-bis (isopropyl)-5-hydroxymethyl-5-amino-hexahydropyrimidine 1,3-bis (B-hydroxyethylaminopropyl)-5-methyl-5-aminohexahydro-pyrimidine 1,3-bis 1, l-dimethyl-Z-hydroxyethyl) -5-methyl-S- amino-hexahydro-pyrimidine 1,3-bis (butylaminopropyl)-5-methyl-5-aminohexahydro-pyrimidine 1,3-bis (ethylhexyl)-5-amino-hexahydro-pyrimidine 1,3-bis (ethylhexyl)-5-hydroxymethyl-5-aminohexahydro-pyrimidine 1,3-bis (cyclohexyl)-5-methyl-S-amino-hexahydropyrimidine 1,3 bis (dicyclohexyl) 5 methyl-S-amino hexahydropyrimidine l,3-bistolyl-S-methyl-5amino-hexahydro-pyrimidine 1,3-bisbenzyl-5-methyl-5-amino-hexahydro-pyrimidine o a 1,3-bis (methylbeiizyl)--methyl-5-amino-hexahydropyrimidine 1,3-bisphenyl-S-methyl-5-amino-hexahydro-pyrimidine 1,3-bis (phenylethyl)-5-methyl-S-amino-hexahydrococcus pyogenes var. aureus and Corynebacterium pyogenes. The novel compositions of this invention find wide application in the treatment of various skin infections due to one or more of the organisms mentioned and pyrimidine 5 are also elfective treatment for various fungus infections 1,3-bis (ethylhexyl)-5-ethyl-5-amino-hexahydroof the scalp or the skin, for certain vaginal infections, as pyrimidine well as for many nose, throat and gingival infections due 1,3-bisdodecyl-S-ethyl-S-amino-hexahydro-pyrimidine to the above and other bacteria and fungi. 1,3-bis (hydroXy tertiary butyl)-5-ethyl-5-an1ino- Any conventional solid or liquid pharmaceutical vehicle hexahydro-pyrimidine or carrier may be employed in preparing these composi- 1,3-bis (cyclohexyl)-S-ethyl-5-amino-hexahydrotions. For topical application, the compositions may be pyrimidine formulated as an aqueous solution, a tincture, an oint- 1,3-bis (ethylhexyl)-5-propyl-5-amino-hexahydroment, a lotion, a salve, a cream, a gel, as a suppository or pyrimidine, and as an aqueous or non-aqueous spray. In dry, powdered 1,3-bis (ethylhexyl)-5-butyl-5-amino-hexah droform these compositions are also quite valuable for the y n pyrimidine treatment of fungus infections. Stable compositions are dily obtained with the usual carriers employed in In lieu of employing the S-ammo-hexahydro-pyrimidlea ines above in the form of their free bases, the salts of gg2fiigiggg 5 222" for the p rep Manon ofthe dosage these amine compounds may also be utilized. Salts are In order further to illustrate our invention but without readily formed with acids such as, for example, acetic beino limited thareto the followino exam la a acid, phosphoric acid, hydrochloric acid, maleic acid, p 5 re g1 benzoic acid, citric acid, malic acid, oxalic acid, tartaric EXAMPLE I i i g acld gilutanc i l i 2 parts by weight of 1,3-bis(beta-ethylhexyl)-5-methyll f fi hff g F ac1d f fg fi S-amino-hexahydro-pyrimidine were dissolved in 1000 su aniliclacif,p itic atilan P- i i f h parts by weight of distilled water to form a 0.2% by j c l qd T? ahp weight aqueous solution. The solution obtained was id sue as t q Pa mmc (new acldmynsnc divided and to one portion of this divided solution was TL h i K ac! h d o 1 added 2.5 parts by weight of ethylenediamine tetraacetic le cdeating agents W fh qd j '5 i g g 3O acid yielding a solution containing 0.2% by weight of com f F y said substituted hexahydro-pyrimidine and 0.5% by pyrimidines are ethylenediamine tetraacetic acid and its weight of ethylenediamine tetraacetic acid sevefal la gf g i m et hy1eneq1am1r 1etet{a The comparative effect of these solutions on the skin actwc (5P t t l l z e1 2 tetlailcetlc 3? flora of the human axilla was determined as follows: te rfiso 1um e Yene mace 1c a F 35 Bacterial samples of the axilla were first obtained as sodlum Calmum ethylenfi'd lamme tetFaaenc acld We a basis for comparison, by rotating a sterile cotton swab, a fourfd that dlethylenetrlamme Pentaacetlc presoaked in di tilled water, over a prescribed skin area, acid and its sodium and calciumsalts are also valuable placing the swab in 10 ml. of gelatin phosphate, and then 1n formulating our novel compositions. Uramil diacetic culturing an aliquot of the sample on nutrient agar under mild fi f of fi d Fllelatmg ageflts which 40 aerobic and anaerobic conditions. The surviving bacterial are sulta f or use t e composlllons of our lrnventlcm' colonies were counted and recorded and this count served Others which may be employed include N,N -ethylene as h base]ine y YP Y y stated, the At zero time the right axilla received a single spray of l n g agents most useful re those which form che- 0.6 gram of the combined solution which is equal to an lates with Mg, Cu, Mn, Co, Fe, Zn and Ca. application of about 228 micrograms of the 1,3-bis(beta- In formulating the novel compositions of our invention ethylhexyl) 5 methyl-S-amino-hexahydro pyrimidine. we preferably employ a suitable pharmaceutical vehicle The left axilla fecel'vfid Seven repealed p y of the or carrier i which h necessary amounts of the active solution (minus the ethylenediamine tetraacetic acid), the components are incorporated to yield a composition conapphcanons being l w to dry between the Separate taming from OJ to 10% by weight of the chelating agent sprays. A total application of about 1596 micrograms of and from 0.01 to 0.5% by weight of the S-amino-hexag l i lethylhexyl) 5 methyl ammo hexa' hydro-pyrimidine. The resulting compositions are highly y m pyrimldme was thus made Bacterial samp were taken at 1, 6 and 24 hours from each axilla and the sameffective on topical application against the organisms Ps.

I ples cultured in the manner described USll'lV BHl broth aerugmosa, P. vulgaris, E. coli and C. albicans and have b en shown t b H ff f 5, and nutrient agar for aerobic culture, and thioglycollate to th f S 0 he eqlua y eLec we in controlling the broth and BBL anaerobic agar for the anaerobic cujture g w 0 ep. emo ytzcus, actobaclllus casez, Micromedium. The results obtained were as follows:

Table I Timeof T fid A bi A of Axilla dili iti ns nu ger t D31210 30212; oii sample B orL I c m h s o lslasil lple colonies colonies aerobic anaerobic e x 10 x it) 0 Right... 5 35 46 3.5 4.5

39 45 Left 5 81 134 9.7 11.8

114 103 1 Right--- 4 3g 35 0.3 0.33 Left 3 is 2% 0.25 0.20 6 Right... 4 156 0.1

Left 4 2% 2.2 24 Right"--- 4 0.1

10 Left 6 107 11.0

5 EXAMPLE n The following components are combined to form a composition suitable for nasal instillation.

Components Parts 1,3 bis (beta ethylhexyl) 5 methyl 5-amin0 hexahydropyrimidine gm 1.0 Polyoxyethylene s o r b it a n mono-oleate (polysorbate 80) gm 14.0 Chlorobutanol gm 5.5 Sodium chloride gm 8.0 Diethylene triamine pentaacetic acid gm 5.0 Sodium h y d r xi d e (10% solution,

w./v.) cc 15.0 Distilled water (q.s.) cc 1000.0

The components are combined in the following manner:

The 1,3-bis(beta-ethylhexyl)--rnethyl-5-amino hexahydropyrimidine and polysorbate 80 are dissolved in 50 parts by weight of distilled water. The chlorobutanol and sodium chloride are separately dissolved in 850 parts by Weight of distilled water heated to 80 C. and the diethylene triamine pentaacetic acid is dissolved in the sodium hydroxide solution. The separate solutions thus formed are combined and sufficient distilled water added to volume.

EXAMPLE III An ointment is prepared from the following components. Components: Parts 1,3 bis (beta ethylhexyl) 5 methyl S-amino hexahydropyrimidine gm 1.0 Polysorbate 80 gm 10.0 Polyethylene glycol (mol. wgt. about 400) gm-.. 500.0 Polyethylene glycol (mol. wgt. about 4000) gm 400.0 Succinic acid grn 0.4 Diethylene triamine pentaacetic acid gm 5 .0 Sodium h y d r 0 xi d e solution,

w./v.) cc 15.0 Distilled water (q.s.) gm 1000.0 To 'form the ointment preparation the 1,3-bis-betaethylhexyl)-5 methyl-5 amino hexahydropyrirnidine is dissolved in the polysorbate 80 and added to a previously melted mixture of the polyethylene glycols. A previously prepared solution of the succinic acid in 50 cc. of Water is added followed by the addition of a solution of the diethylene triamine pentaacetic acid in the sodium hydroxide solution. After thorough mixing, the composition is adjusted to Weight by the addition of water.

EXAMPLE IV A shampoo preparation is formulated from the follow ing components in the manner described.

Components: Parts 1,3 bis (beta ethylhexyl) 5 methyl S-amino hexahydropyrimidine gm 1.0 Polyethylene glycol (mol. wgt. about 400) distearate gm 16.7 Ammonium lauryl sulfate (27% solution) I gm 500.0 'Diethylene triamine pentaacetic acid gm 5.0 Sodium h y d r o x i d e (10% solution,

-w./v.) cc 15.0 Distilled water cc 20.0 Ethyl alcohol (q.s.) cc 1000.0 Perfume (optional) cc There are then added, in order, the remainder of the amone liter with alcohol and the perfume added.

EXAMPLE V A colloidal suspension suitable for therapeutic appli- Cation is formed by combining the following components.

Components: Parts 11,3 bis (beta ethylhexyl) 5 methyl 5-amino hexahydropyrimidine grn 1.0 Hydrochloric acid, 10% (q.s., pH 2.0-2.5). Polyvinylpyrrolidone gm 40.0 Sodium hydroxide, 30% solution (pH 6.8-

7.2). Sodium carboxymethyl cellulose (Hercules HV) grn 20.0 Diethylene triamine pentaacetic acid gm 50.0 Sodium 11 y d r 0 xi d e (10% solution,

w./v.) cc 150.0 Distilled water (q.s.) cc 1000.0

In forming the suspension, the hydrochloric acid is added to the l,3-bis(beta-ethylhexyl) -5-methyl-5-aminohexahydropyrimidine until a pH of 2.0-2.5 is reached, a solution of the polyvinylpyrrolidone in 40 cc. of distilled water is then added and the pH is adjusted to 6.87.2 with the sodium hydroxide solution. A previously prepared solution of the sodium carboxymethyl cellulose in 600 cc. of distilled water is added followed by a solution of the diethylene triamine pentaacetic acid in the 10% aqueous sodium hydroxide solution. The preparation is thoroughly mixed and adjusted to the indicated volume With distilled water.

EXAMPLE VI A colloidal suspension is formed of the following.

Components: Parts 1,3-bis (beta-ethylhexyl)-5-methyl-5-aminohexahydro-pyrimidine "gm-.. 5.0 Hydrochloric acid (10% aqueous solution) q.s. Polyvinylpyrrolidone gm 40.0 Sodium hydroxide (30% aqueous solution)- q.s. Sodium carboxymethyl cellulose (Hercules 70 HV) grn 16.6 Polyethylene glycol (mol. wgt. about 400) gm 41.6 Sodium chloride gmu 4.0 Sodium benzoate gm 1.5 Ethylene diamine tetraacetic acid gm 100.0 Distilled water (q.s.) grn 1000.0

The hydrochloric acid solution is added to the 1,3-bis- (beta-ethylhexyl)5-amino-hexahydro-pyrimidine until a pH of 2.0-2.5 is reached. To this mixture is then added a solution of the polyvinylpyrrolidone in 40 cc. of distilled Water and the pH then adjusted to 6.8-7.2 by adding a sufficient amount of the sodium hydroxide solution. This mixture is then added to a previously prepared solution of the sodium carboxymethyl cellulose in 500 cc. of distilled Water to which the polyethylene glycol, sodium chloride, and sodium benzoate have been added. To form the final suspension a solution of the ethylene diamine tetraacetic acid in sodium hydroxide solution is added, the

volume is adjusted and the suspension formed is thoroughly mixed.

7 EXAMPLE VII A stable emulsion is prepared from the following.

Components: Parts 1,3-bis-(beta-ethylhexyl)--rnethyl-5-aminohexahydro-primidine gm 4.00 Brominated vegetable oil gm 4.00 Methyl parahydroxybenzoate gm- 0.80 Propyl parahydroxybenzoate gm 0.16 Polysorbate 80 gm 8.00 Methyl cellulose (400 cps. viscosity) gm 8.00 Glyceryl monostearate, pure gm 28.00 Diethylene triamine pentaacetic acid grn 50.00

Sodium hydroxide solution w./v.)

cc 150.00 Distilled water (q.s.) cc.. 1000.00

The glyceryl monostearate is heated to 70 C. and a mixture of the 1,3-bis-(beta-ethylhexyl)-5-methyl-5- amino-hexahydropyrimidine, brominated vegetable oil, and polysorbate 80 previously heated to 70 C. is added. The propyl parahydroxybenzoate i then dissolved in this melt and the mixture added to a solution of the methyl parahydroxybenzoate in 400 cc. of distilled water heated to 70 C. to which had been added the diethylene triamine pentaacetic acid in the aqueous sodium hydroxide solution. Stirring is continued while heating this portion of the batch to 90 C. and then stirring is continued until cooled to room temperature. The methyl cellulose is soaked in 125 cc. of distilled water at 90 C., mixed for about 30 minutes and then 125 cc. of distilled Water, cooled to 4 C. is added slowly while mixing. The mixture obtained is stored at refrigerator temperature over night and, after the temperature is raised to room tempera ture, the latter is added to the 1,3-bis(beta-ethylhexyl)- 5-methyl-5-amino-hexahydro-pyrimidine solution previously formed, the whole mixed well, and then adjusted to volume with distilled water.

EXAMPLE VIII A suitable therapeutic ointment is formed from the following.

The 1,3-bis-(beta-ethylhexyl)-5-methyl-5-amino hexahydropyrimidine is dissolved in the polysorbate 80 and added to a previously melted mixture of the polyethylene glycol. A previously prepared solution of the succinic acid in 50 cc. of water is then added. The uramil diacetic acid is triturated with about 10 gm. of the base material, the latter then mixed with the remainder of the base and the ointment formed is finally adjusted to weight with distilled water.

It is understood that the foregoing detailed description is given merely by way of illustration and that many variations may be made therein without departing from the spirit of my invention.

Having described our invention, what we desire to secure by Letters Patent is:

What we claim is:

1. A therapeutic composition comprising a pharmaceutical carrier, from 0.1 to 10% by Weight of a chelating agent selected from the group consisting of ethylenediamine tetraacetic acid, diethylenetriamine pentaacetic acid, uramil diacetic acid, N,N-ethylene bis [Z-(Z-hydroxyphenyl)] glycine and salts thereof and from 0.01 to wherein R is a radical of the group consisting of alkyl, aryl, aralkyl, alkaryl, hydroxyalkyl, aminoalkyl and cycloalkyl radicals and R is selected from the group consisting of hydrogen, lower alkyl and hydroxymethyl, and the non-toxic salts of said 5-amino-hexahydro-pyrimidine.

2. A therapeutic composition as defined in claim 1 wherein the chelating agent is ethylenediamine tetraacetic acid.

3. A therapeutic composition as defined in claim 1, wherein the chelating agent is diethylenetriamine pentaacetic acid.

4. A therapeutic composition as defined in claim 1, wherein the chelating agent is uramil diacetic acid.

5. A therapeutic composition as defined in claim 1 wherein the chelating agent is disodium ethylenediamine tetraacetic acid.

6. A therapeutic composition as defined in claim 1, wherein the chelating agent is N,N -ethylene bis-[2-(O- hydroxyphenyl) glycine.

7. A therapeutic composition comprising a pharmaceutical carrier, from 0.1 to 10% by Weight of ethylenediamine tetraacetic acid and from 0.01 to 0.5% by weight of a member of the group consisting of 1,3-bis (beta-ethylhexyl)-5-methyl-5-amino hexahydro pyrimidine, and the non-toxic salts thereof.

8. A therapeutic composition comprising a pharmaceutical carrier, from 0.1 to 10% by weight of diethylenetriamine pentaacetic acid and from 0.01 to 0.5 by Weight of a member of the group consisting of 1,3-bis (beta-ethylhexyl)-5-methyl-5-amino-hexahydro-pyrimidine, and the non-toxic salts thereof.

9. A therapeutic composition comprising a pharmaceutical carrier, from 0.1 to 10% by weight of uramil diacetic acid and from 0.01 to 0.5% by weight of a member of the group consisting of 1,3-bis(beta-ethylhexyl)-5- methyl-5-amino-hexahydro-pyrimidine, and the non-toxic salts thereof.

10. A therapeutic composition comprising a pharmaceutical carrier, from 0.1 to 10% by weight of N,N -ethylene bis-[Z-(O-hydroxyphenyl)] glycine and from 0.01 to 0.5% by Weight of a member of the group consisting of 1,3-bis(beta-ethylhexyl)-5-methyl-5 amino hexahydropyrimidine, and the non-toxic salts thereof.

11. A therapeutic composition comprising a pharmaceutical carrier, from 0.1 to 10% by weight of disodium ethylenediamine tetraacetic acid and from 0.01 to 0.5% by Weight of a member of the group consisting of 1,3-bis (beta-ethylhexyl)-4-methyl-5-amino hexahydro pyrimidine, and the non-toxic salts thereof.

References Cited in the file of this patent UNITED STATES PATENTS Fosdick June 3, 1958 OTHER REFERENCES 

1. A THERAPEUTIC COMPOSITION COMPRISING A PHARMACEUTICAL CARRIER, FROM 0.1 TO 10% BY WEIGHT OF A CHELATING AGENT SELECTED FROM THE GROUP CONSISTING OF ETHYLENEDIAMINE TETRAACETIC ACID, DIETHYLENETRIAMINE PENTAACETIC ACID, URAMIL DIACETIC ACID, N,N''-ETHYLENE BIS 2-(2-HYDROXYPHENYL)! GLYCINE AND SALTS THEREOF AND FROM 0.01 TO 0.5% BY WEIGHT OF A MEMBER OF THE GROUP CONSISTING OF A 5-AMINO-HEXAHYDRO-PYRIMIDINE OF THE FORMULA 